Automated EuroFlow approach for standardized in-depth dissection of human circulating B-cells and plasma cells

Frontiers in Immunology, Год журнала: 2023, Номер 14

Опубликована: Окт. 17, 2023

Multiparameter flow cytometry (FC) immunophenotyping is a key tool for detailed identification and characterization of human blood leucocytes, including B-lymphocytes plasma cells (PC). However, currently used conventional data analysis strategies require extensive expertise, are time consuming, show limited reproducibility.Here, we designed, constructed validated an automated database-guided gating (AGI) approach fast standardized in-depth dissection B-lymphocyte PC populations in blood.For this purpose, 213 FC standard (FCS) datafiles corresponding to umbilical cord peripheral samples from healthy patient volunteers, stained with the 14-color 18-antibody EuroFlow BIgH-IMM panel, were used.The antibody panel allowed 117 different subsets. Samples 36 donors 14 that fulfilled strict inclusion criteria analysed by expert cytometrist build database. Data contained was then merged into reference database uploaded Infinicyt software (Cytognos, Salamanca, Spain). Subsequently, compared results manual (MG) performance two classification algorithms -hierarchical algorithm vs two-step algorithm- AGI cell present 5 randomly selected FCS datafiles. The hierarchical showed higher correlation values MG (r2 0.94 vs. 0.88 algorithm) further set 177 against expert-based MG. For virtually all identifiable highly significant observed between approaches (r2>0.81 79% B-cell identified), significantly lower median per sample (6 40 min, p=0.001) MG, respectively both intra-sample (median CV 1.7% 10.4% p<0.001) inter-expert 3.9% 17.3% 2 experts, variability.Our strategies, here proposed faster, more robust, reproducible, subsets circulating blood.

Язык: Английский

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Altered immune cell phenotypes within chronically ischemic human kidneys distal to occlusive renal artery disease

AJP Renal Physiology, Год журнала: 2023, Номер 326(2), С. F257 - F264

Опубликована: Ноя. 30, 2023

Renal artery stenosis (RAS) is a major cause of ischemic kidney disease, which largely mediated by inflammation. Mapping the immune cell composition in kidneys might provide useful insight into disease pathogenesis and uncover therapeutic targets. We used mass cytometry (CyTOF) to explore single-cell unique data set human nephrectomized due chronic occlusive vascular (RAS,

Язык: Английский

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Comparative assessment of cytometry by time-of-flight and full spectral flow cytometry based on a 33-color antibody panel

Journal of Immunological Methods, Год журнала: 2024, Номер 527, С. 113641 - 113641

Опубликована: Фев. 15, 2024

Mass cytometry and full spectrum flow have recently emerged as new promising single cell proteomic analysis tools that can be exploited to decipher the extensive diversity of immune repertoires their implication in human diseases. In this study, we evaluated performance mass against using an identical 33-color antibody panel on four healthy individuals. Our data revealed overall high concordance quantification major populations between two platforms a semi-automated clustering approach. We further showed strong correlation cluster assignment when comparing manual automated clustering. Both comparisons minor disagreements rare subpopulations. study both technologies generate highly overlapping results substantiate choice technology is not primary factor for successful biological assessment profiles but must considered broader design framework clinical studies.

Язык: Английский

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Application of Mass Cytometry Platforms to Solid Organ Transplantation

Transplantation, Год журнала: 2024, Номер 108(10), С. 2034 - 2044

Опубликована: Март 12, 2024

Transplantation serves as the cornerstone of treatment for patients with end-stage organ disease. The prevalence complications, such allograft rejection, infection, and malignancies, underscores need to dissect complex interactions immune system at single-cell level. In this review, we discuss studies using mass cytometry or by time-of-flight, a cutting-edge technology enabling characterization populations cell-to-cell in granular detail. We review application human experimental animal context transplantation, uncovering invaluable contributions tool understanding rejection other transplant-related complications. recent innovations that have potential streamline standardize workflows multisite clinical trials. Additionally, introduce imaging cytometry, technique couples power spatial context, thereby mapping cellular within tissue microenvironments. synergistic integration data omics sets high-dimensional platforms further define dynamics is discussed. conclusion, technologies, when integrated tools data, shed light on intricate landscape response transplantation. This approach holds significant enhancing patient outcomes advancing our facilitating development new diagnostics therapeutics.

Язык: Английский

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The over-activated peritoneal immune environment in endometriosis is characterised by a lack of PD-1 inhibition.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 2, 2024

Abstract Background Endometriosis is characterised by chronic inflammation in the peritoneal cavity causing acute and pelvic pain, largely explained dysregulation immune environment within fluid. The activation status of cells unclear. In addition, a comparison with systemic system desirable to explore avenues diagnosis treatment endometriosis-related pain. Objective(s) To investigate endometriosis fluid blood full-spectrum flow cytometry focus on inhibition cells. Study Design This was an observational study patients undergoing laparoscopy for or unrelated conditions; PF collected from n=5 n=4 controls, n=3 controls. Data were analysed statistical significance using ANOVA, Kruskal-Wallis Mann-Whitney U test, p-value below 0.05 considered significant. Results We observed prevailing myeloid as opposed lymphoid blood. main differences between control samples, however, found smaller compartments, i.e., populations PD-1 levels samples significantly lower than controls (p<0.05). Conclusion(s) checkpoint could be new angle treating pain women suffering this intractable condition. Tweetable Lack plays role inflammation. AJOG at glance inflammatoru needs investigating it iis causative predominant symptom. lack expression compared explain persistent open treatment.

Язык: Английский

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